The following antibodies and reagents were used in this study: anti-phospho-JNK (Cell Signaling 4668); anti-phospho-P38 (Cell Signaling 4511); anti-phospho-ERK (Cell Signaling 4370); anti-phospho-NF-κBp65 (Cell Signaling 3033); anti-phospho-NF-κBp105 (Cell Signaling 3035); anti-phospho-AKT (Ser473; Cell Signaling 4060); anti-JNK (bioworld BS3630); anti-P38 (bioworld BS3567); anti-ERK (bioworld BS1112); anti-NF-κBp65 (Cell Signaling 4764); anti-AKT (bioworld MB0052); anti-TLR2(Cell Signaling 12276); anti-TLR3 (GeneTex GTX113022); anti-TLR4 (GeneTex GTX125909); anti-A20/TNFAIP3 (Cell Signaling 5630); anti-gC1qR/p33 (Millipore MAB1161); Human C1q were obtained from Sigma (C1740); Inhibitors specific for P38 (SB203580), JNK (SP600125), ERK (PD98059), IKKβ (IMD 0354), PI3K (Wortmaninn) and AKT (MK 2206 2HCl) were obtained from selleckchem. CBA Kit was obtained from BD. Ni-NTA Agarose were obtained from QIAGEN. His peptide was obtained from Chinese Peptide Company.
Anti erk
Manufactured by Bioworld Technology
Sourced in United States, China
Anti-ERK is a laboratory reagent used in biological research. It is a monoclonal antibody that specifically binds to the extracellular signal-regulated kinase (ERK) protein, which is a key component of the MAPK signaling pathway. Anti-ERK can be used in various research applications, such as Western blotting, immunoprecipitation, and immunohistochemistry, to detect and quantify ERK expression and activation.
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Lab products found in correlation
Anti p erk, by Bioworld Technology (5 mentions)
Anti akt, by Bioworld Technology (4 mentions)
Anti jnk, by Bioworld Technology (3 mentions)
Anti p akt, by Bioworld Technology (3 mentions)
Anti p38, by Bioworld Technology (2 mentions)
Cba kit, by BD (1 mentions)
Anti phospho akt ser473, by Cell Signaling Technology (1 mentions)
Anti tlr2, by Cell Signaling Technology (1 mentions)
Anti phospho nf κbp105, by Cell Signaling Technology (1 mentions)
Anti phospho erk, by Cell Signaling Technology (1 mentions)
Human c1q, by Merck Group (1 mentions)
Anti tlr4, by GeneTex (1 mentions)
Anti phospho nf κb p65, by Cell Signaling Technology (1 mentions)
Pd98059, by Selleck Chemicals (1 mentions)
Sb203580, by Selleck Chemicals (1 mentions)
Anti nf κb p65, by Cell Signaling Technology (1 mentions)
Anti phospho jnk, by Cell Signaling Technology (1 mentions)
Anti phospho p38, by Cell Signaling Technology (1 mentions)
Mk 2206 2hcl, by Selleck Chemicals (1 mentions)
Ni nta agarose, by Qiagen (1 mentions)
6 protocols using anti erk
1
Signaling Pathway Activation Analysis
2
Western Blot Analysis of Spinal Cord Proteins
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Spinal cords were homogenized with a micro content motor-operated tissue homogenizer (Kimble Kontes, USA), using protein extraction kit (Sigma, USA) supplemented with a cocktail of protease inhibitors. The homogenates were centrifuged at 13,000g for 20 min at 4 ℃, and the supernatants were collected. The protein concentration was determined by BCA method. Equal amounts of protein (30 μg) were separated by SDS-PAGE and electroblotted onto PVDF membrane (Immobilon-P, Millipore, USA). After non-specific binding was blocked with 5% non-fat dry milk, membranes were incubated at 4 ℃ overnight with anti-MBP, anti-Akt, anti-ERK, anti-JNK, anti-P-Akt, anti-P-ERK, anti-P-JNK and anti-β-actin (1:1000, Bioworld, USA). After washing the membrane the next day, it should be incubated with the second antibody at room temperature for 2 hours. Bands were visualized with an enhanced chemiluminescence (ECL) system (GE Healthcare Life Sciences, USA).
3
Protein Extraction and Western Blot Analysis in MCF-7 Cells
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MCF-7 cells were harvested by the use of ice-cold RIPA lysis buffer (Beyotime Biotechnology, Shanghai, China) and protein concentration was determined using the BCA protein quantification kit (Beyotime Biotechnology, Shanghai, China); western blotting was performed as previously described [19 (link)]. Primary antibodies of anti-Ac-H3, anti-SOD1, anti-CAT, anti-ERK, anti-p-ERK, anti-Cyt-C, and anti-caspase 9 were purchased from Bioworld Technology (Nanjing, China), anti-PARP, rabbit anti-caspase 3, and anti-Ac-H3 were purchased from Cell Signaling Technology, Danvers, USA, anti-RAS was purchased from BD, USA, and anti-GAPDH was purchased from Earthox, Millbrae, USA. The secondary antibodies were purchased from Biogot Biotechnology (Nanjing, China), and ECL SuperSignal West Femto Maximum Sensitivity Substrate was purchased from Thermo Fisher.
4
Western Blot Analysis of Protein Expression
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We carried out western blot as previously described [33 (link)], using anti-TLE3 (Abcam, Cambridge, MA, USA), anti-FOXO3, anti-Akt, anti-GSK-3β, anti-ERK, anti-p21, anti-p27, anti-p-FOXO3, anti-p-Akt, anti-p-GSK-3β and anti-p-ERK (Bioworld Technology, St. Louis Park, MN, USA) to detect the corresponding proteins. Anti-α-Tubulin monoclonal antibody (Sigma, St. Louis, MO, USA) served as a loading control.
5
Endothelial Cell Migration and Proliferation
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Fetal bovine serum, Trizol, Lipofectamine 2000and HRP-conjugated mouse and rabbit secondary antibodies were obtained from Invitrogen (Shanghai, China). Endothelial Cell Medium (ECM) was obtained from ScienCell (San Diego, USA). The anti-AXIIR and anti-CD31antibody were obtained from Santa Cruz Biotechnology Co., Ltd. (Dallas, USA). Antibodies to MMP2, MMP9 were obtained from Shanghai RuianBioTechnologies (Shanghai, China). The anti-AKT, anti-p-AKT, anti-ERK, anti-p-ERK, anti-TIMP2 antibodies were purchased from Bioworld Technology, Inc. (Nanjing, China). The anti-GAPDH antibody was obtained from Tianjin sungene Biotech (Tianjin, China). Matrigel Matrix was obtained from BD Biosciences (San Jose, USA). Cell-light EdU DNA cell proliferation kit was obtained from RiboBio (Guangzhou, China). PrimeScript RT reagents Kit, PrimeSTAR® HS DNA Polymerase with GC Buffer and SYBR Premix Ex Taq were obtained from TAKARA BIOTECHNOLOGY (Dalian, China). Annexin V-FITC Apoptosis Detection Kit was obtained from eBioscience (San Diego, CA, USA). The transwell chamber and other cell culture plates were obtained from Corning Inc (NY, USA). All other regents were of analytical grade.
6
Protein Expression Analysis in Melanoma
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Melanoma tissues or A375 cells were lysed in radioimmunoprecipitation assay lysis buffer (Beyotime Institute of Biotechnology, Haimen, China), and the protein concentrations of the cell lysates were determined using the Bicinchoninic Acid protein assay kit (Beyotime Institute of Biotechnology). Lysates (30 µg) were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and then transferred to polyvinylidene fluoride membranes. The membranes were blocked with 5% milk and then probed at 4°C overnight with the following primary antibodies: anti-SPAG9, anti-E-cadherin, and anti-vimentin (1:2500, Bioworld Technology, St. Louis Park, MN, USA); anti-MMP2, anti-MMP9, and anti-GAPDH (1:3500, Bioworld Technology); and anti-p38, anti-p-p38, anti-ERK, anti-p-ERK, anti-JNK, and anti-p-JNK (1:4500, Bioworld Technology). The membranes were then incubated with horseradish peroxidaselabeled goat anti-rabbit IgG (1:5000, Bioworld Technology) for 2 h, and immunoreactivity was assessed using enhanced chemiluminescence detection reagent (Beyotime Institute of Biotechnology) and ImageJ software.
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